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    • 4. 发明申请
    • METHODS AND REAGENTS FOR THE DETERMINATION OF IMMUNOSUPPRESSIVE AGENTS
    • 用于确定免疫抑制剂的方法和试剂
    • WO1993025533A1
    • 1993-12-23
    • PCT/US1993005197
    • 1993-06-01
    • ABBOTT LABORATORIES
    • ABBOTT LABORATORIESLANE, Benjamin, ClayLULY, Jay, RichardSMITH, Allan, H.BOLLING, Timothy, J.MANDECKI, WldozimierzPILOT-MATIAS, Tami, J.
    • C07D221/18
    • C07D498/18C07K14/715C07K2319/32C07K2319/61C12N9/1241
    • Assay methods and reagents for determining the presence or amount of immunophilin ligands and immunophilins thereof employing a recombinant fusion protein comprising (i) an immunosuppressant binding protein and (ii) a heterologous protein are disclosed. The recombinant fusion protein can also be employed for the evaluation of immunosuppressive activities of immunosuppressive agents in order to determine the efficacy of an immunosuppressive agent during the course of therapeutic treatment of a patient therewith. Preferably, the recombinant fusion protein comprises a macrolide immunosuppressive agent and CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyl transferase. When employed in a binding assay format, the recombinant fusion protein provides higher reactivity for the immunophilin ligand under determination than does the native immunosuppressant binding protein. In particular, an immunosuppressant assay reagent comprising FK-506 binding protein (FKBP) and CKS immobilized to a solid support material provides a higher signal-to-noise ratio when employed in a competitive heterogeneous assay format than when native FKBP immobilized to a solid support material is employed in such assay format.
    • 公开了使用包含(i)免疫抑制剂结合蛋白和(ii)异源蛋白质)的重组融合蛋白来测定免疫亲和素配体及其免疫亲和物的存在或量的测定方法和试剂。 重组融合蛋白也可以用于评估免疫抑制剂的免疫抑制活性,以确定免疫抑制剂在其患者治疗过程中的功效。 优选地,重组融合蛋白包含大环内酯免疫抑制剂和CTP:CMP-3-脱氧-D-壬烯 - 辛酸酯胞苷酰转移酶。 当以结合测定形式使用时,重组融合蛋白质在测定时比天然免疫抑制剂结合蛋白提供更高的免疫亲和素配体的反应性。 特别地,当以竞争性异质测定格式使用时,固定在固体支持物质上的包含FK-506结合蛋白(FKBP)和CKS的免疫抑制剂测定试剂提供比当固定在固体支持物上的天然FKBP时更高的信噪比 材料以这种测定形式使用。